IGEM:Peking/2007/Switch-Notebook/2007-7-11

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Experimenter: LCB, JLL

Contents

Contents

Precipitate with ethanol: pLX007 (X/S)

  • Time: 8:30-12:30

Temperature PCR of SD2 (X/S)

  • Template: SD2-pMD18T
  • Primers: lacO2_XhoI_XbaI_F, lacO2_SalI_SpeI_R

(Woooo.. I forgot to take the picture..)

Failed.

Colony PCR test of SDD’-EGFP-pCC002, RD1, CD1, SS1-3, SD3-pMD18T

Image:SS1 SD3 pcr test larger.gif

l to r: Negative control, SS1(1-10), SD3(13-18), SS3(25-30), RD1, Marker

Image:SS3 SS2 SDD'-EGFP-pCC002 pcr test.gif

l to r: SS3(31-32), SS2(37-44), SDD'-EGFP-pCC002(1-12), none, Negative control for SS2, Marker

Image:PKU Switch 07-7-11 SDD'-EGFP-pCC002 SDD'-EGFP-BamHI-F&pCCtestR.gif

SDD'-EGFP-pCC002(13-35), Negative Control, Marker

Send for sequencing: bla…bla…bla…

收ECFP-pCC002新连的板子

自连&连接长得差不多!

Ligation of pLX007 (X/S) and lacZa

Ordered primers arrived

  • SDA-H, cro, Y_lacZa_NotI_F

PCR test of CD1(1-6), RD1(1-6)-pMD18T

Photo omitted. The result is not good. We need to repeat this.

PCR of various SD_lacZa

Need to be repeated.

Colony PCR test of pLX007-lacZa

  • testF&R

Failed..

Ligation and transformation

  • lacZa and pLX007, 4:1 or 4.7:0.3
  • positive transformation of pCC002-lacZa into JM109 and Math-T1

Sequencing result received

SD1-3, SD2-1, SD2-2 are right.

Miniprep and digestion test with BamHI.

Image:PKU Switch 07-7-11 SD1-3 ECFP pCC002.gif


Looking forward

Precipitate pLX007 (S/X)

Check the plates of lacZa-pLX007

Pick whichever is better.

收pCC002-lacZa板子到4C

Send for sequencing

CD1-1, CD1-3, RD1-1

PCR

Various lacZa, pCC002-lacZa

Miniprep of SDD’-EGFP-pCC002(1-8), digestion test

Check the plates of SDD-EGFP-pCC002

收好CD1, RD1, SD3,SS1-3的菌液

Recheck the plates of CD1, RD1-pMD18T

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